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Isolation and characterization of the normal canine β‐galactosidase gene and its mutation in a dog model of GM1‐gangliosidosis
Author(s) -
Wang Z. H.,
Zeng B.,
Shibuya H.,
Johnson G. S.,
Alroy J.,
Pastores G. M.,
Raghavan S.,
Kolodny E. H.
Publication year - 2000
Publication title -
journal of inherited metabolic disease
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.462
H-Index - 102
eISSN - 1573-2665
pISSN - 0141-8955
DOI - 10.1023/a:1005630013448
Subject(s) - biology , microbiology and biotechnology , peptide sequence , nucleic acid sequence , amino acid , gene , complementary dna , mutation , exon , point mutation , biochemistry , genetics
The acid β‐galactosidase cDNA of Portuguese Water dogs was isolated and sequenced. The entire coding region of the gene consists of 2004 nucleotides encoding a protein of 668 amino acids. Its encoding sequence indicates approximately 86.5% identity at the nucleotide level and about 81% identity at the amino acid level with the encoding region of the human acid β‐galactosidase gene. The deduced amino acid sequence contains a 24‐amino‐acid putative signal sequence, six possible glycosylation sites, and seven cysteine residues. A homozygous recessive mutation, causing canine GM1‐gangliosidosis, was identified at nucleotide G 200 →A in exon 2 resulting in an Arg 60 →His (mutation R60H) amino acid substitution. The mutation creates a new restriction enzyme site for Pml 1. Genotyping 115 dog samples for this acid β‐galactosidase gene alteration readily distinguished affected homozygous recessives ( n =5), heterozygous carriers ( n =50) and normal homozygotes ( n =60). DNA mutation analysis provided a method more specific than enzyme assay of β‐galactosidase for determination of carriers.